Enzyme-linked immunosorbent assay for products of the 60-megadalton plasmid of Escherichia coli serotype O157:H7.

Abstract Eighty strains of pathogenic Escherichia coli, representing each of the major diarrheal disease-causing groups, were examined by direct enzyme-linked immunosorbent assay (ELISA) for the presence of proteins associated with a 60-MDa plasmid from E. coli serotype O157:H7. Antiserum specific for plasmid-encoded proteins was prepared by immunizing a rabbit with a wild-type E. coli O157:H7 strain (strain 7785) and absorbing the serum with a plasmid-cured derivative (strain 2-45). Use of this antiserum in Western immunoblot analysis detected two proteins of 82 and 92 kDa in strain 7785 but not in strain 2-45. All 16 wild-type E. coli O157:H7 strains and all 10 Shiga-like toxin (SLT)-producing E. coli strains of serotypes other than O157 were ELISA positive. Thirteen of 14 enterotoxigenic and all of 24 enteroinvasive E. coli strains were ELISA negative, as were all of 16 E. coli strains isolated from healthy persons. Of 16 traditional enteropathogenic E. coli (EPEC) serotypes, 10 were ELISA positive, including 10 of 12 strains carrying the EPEC adherence factor gene. Absorption of the serum with an EPEC adherence factor-positive EPEC eliminated EPEC reactivity. This study demonstrates that two plasmid-mediated proteins are common to E. coli O157:H7 strains and to SLT-producing strains of other serotypes. Detection of these proteins by ELISA provides a sensitive and specific screening test for identifying SLT-producing E. coli of both O157 and non-O157 serotypes. Identification of the cross-reactive proteins found in EPEC could provide the basis for a single assay to detect both EPEC and SLT-producing E. coli.